Apparatus and method for the purification of biomolecules

ABSTRACT

The invention comprises an apparatus  10  and a method  100  for automatically lysing, extracting and purifying biomolecules  28 . As the central technology for purifying the biomolecules magnetizable particles  35  and an adapted magnet device  40  are used for binding and transferring the particles  35 . The apparatus  10  comprises a highly efficient incubation unit  60  with options for upscaling the maximum sample amount. Furthermore the apparatus  10  in particular comprises a group of magnetizable pins  40  as transport magnets and furthermore counter magnets  50  which are arranged on the cavities  20 , in which solutions  30  with the biomolecules  28  to be lysed and magnetizable particles  35  are disposed. The use of counter magnets  50  improves the quality of the eluate. Furthermore the invention comprises a fully automatic system  800  and a method  100  for controlling the process steps and the selection of the reagents and the aids (e.g. magnetizable particles  35 ) on the basis of automatically detected information, such as gathered e.g. from the loaded samples and/or from the used reagents. The automated overall system  800  in particular fulfils the requirements of medical diagnostics.

This application claims the priority from the German patent application DE 10 2007 054 033.9, to which reference is made and which in its entirety forms part of this disclosure. This application is filed concurrently in Germany and the USA.

FIELD OF THE INVENTION

The invention relates to a modular apparatus and a composite multistage method as well as a comprehensive system for the automated lysis, extraction and purifying of biomolecules (e.g. DNA, RNA or proteins). The method steps according to the invention comprise recognizing substances, placing the substances in cavities, lysis, binding or adsorbing of biomolecules to magnetizable particles, transferring particles, washing and cleaning of biomolecules, eluting or desorbing the biomolecules, removing the magnetizable particles and pipetting the eluate into a yield cavity. The system fulfils the requirements of clinical in-vitro diagnostics.

DESCRIPTION OF RELATED ART AND BACKGROUND OF THE INVENTION

There are a very large number of suppliers of apparatus and methods for the purifying of biomolecules via adsorption to magnetizable particles. The known methods are determined by the type of biomolecules and their interaction with the magnetizable particles. These types of purifying methods are in particular adapted for the automation of the methods for purifying of biomolecules.

Among the suppliers of apparatus and methods for the purifying of biomolecules the following suppliers are known: Chemagen, Qiagen, Thermo, Promega, Roche Diagnostics, Agowa, Dynal, Thermo Fisher, Analytik Jena and Tecan.

These companies partly protected their apparatus through their own patents (for example WO 03/044537, WO 2007/020294, U.S. Pat. No. 6,448,092 and WO9609550).

Most of these known apparatus have in common that they are so-called magnet separators without pipetting function (Thermo Kingfisher, Promega, Agowa). This means an operator pipettes solutions prior to the extraction process by hand or he uses pre-filled plastic cavities for the extraction. In an apparatus supplied by Chemagen the cavities are also filled, but the eluate still needs to be removed by pipetting manually. These approaches are sufficient for applications in a field of research, however, they are not sufficient in the diagnostic field.

An apparatus offering a further solution is the Magnapure technology by Roche Diagnostics.

The European patent application No. 0 644 425 by Hoffmann-La Roche discloses an analysis apparatus with a device for separating magnetic microparticles from a suspension. The apparatus disclosed in the Hoffmann-La Roche application comprises two permanent magnets between which a reaction cavity containing a suspension is disposed. The permanent magnets are arranged diametrically opposite each other with respect to the reaction cavity. The polar axes of the permanent magnets and the longitudinal axis of the reaction cavity form an acute angle. With this arrangement of the polar axes the magnetic stray field can be used for the separation of the magnetic microparticles, thereby accelerating the separation of the magnetic microparticles.

U.S. Pat. No. 6,596,162 (Thermo Labsystems Oy) and U.S. Pat. No. 6,040,192 (Labsystems Oy) disclose an apparatus for separating magnetic particles from a reaction cavity with a removable permanent magnet.

The use of permanent magnets and suspensions with magnetizable particles is the central technology of the known apparatus. Due to the effect of the magnetic force the magnetizable particles are collected from the suspension and adhere to the magnet. After removing the magnetic field these particles are released again. This object is solved in different ways in the known apparatus.

U.S. Pat. No. 6,409,925 (Bio-Magnetics Ltd.) discloses an apparatus for the collecting of magnetic particles as well as the transfer of the magnetic particles from a first cavity to a second cavity. For collecting and transferring the magnetic particles this disclosure suggests the use of a mobile magnetic element in order to magnetize a tip. A magnetization of the tip occurs upon approaching the mobile magnetic element to the tip. The magnetized tip is adapted for collecting and the transferring the magnetic particles.

EP 1 726 963 A2 (Festo Corporation) discloses a system for transferring a sample material from a source cavity to a target cavity. A transfer unit comprises at least one pin tip with a central bore and a magnetic actuator element being moveable between a first and a second actuator position. The sample material in proximity of the pin tip is either collected or released by moving the actuator element. Furthermore a group of counter magnets is provided underneath the source cavity and/or the target cavity, in order to support the transfer of the sample material. The Festo system furthermore allows a common movement of the actuator element and the group of counter magnets in order to mix the sample material. It is furthermore provided that each pin tip is individually controllable; likewise, the movement of each of the counter magnets is individually controllable.

The above-mentioned solutions have the disadvantage of comprising qualitative shortcomings in the production of the eluate; and that their majority does not fulfill the requirements of in-vitro diagnostics, comprehensively.

The purifying of biomolecules using the magnetizable particles leads to “residual particles” in the eluate according to the methods known in the state of the art. These have to be removed at a great effort in a further process step, for example by centrifugation, since they impair subsequent analytic processes.

Furthermore, the concatenation of the process steps from the lysis of the sample to the selection of the substances and aids, via the numerous processing steps yielding the final eluate so far comprised numerous sources of errors typically leading to contamination, falsely negative or falsely positive results.

Furthermore, there is the danger of confusion when combining substances within a kit used for the so-called purifying. A kit consists of a defined combination of substances and aids. The substances and aids comprise e.g. elution buffers. A use of e.g. wrong elution buffers would lead to the biomolecules no longer being separable from the biomolecules bound to the magnetizable particles in the elution buffer, and would thus erroneously no longer be detectable in the eluate. It is essential to prevent this.

The present invention uses among others kits of substances for the purifying of nucleic acids which are available for example from Invitek (disclosed e.g. in the patents: WO01/70386, WO2004/042058, DE10253351, WO00/34463, WO99/32616, U.S. Pat. No. 6,037,465, DE59610721, DE4422044 and others). Specifically, the invention uses magnetizable particles adapted to temporarily bind the biomolecules, thus rendering them transportable.

These kits of substances by Invitek require, in particular during the lysis, temperatures of up to 90° Celsius over a period of for e.g. 20 minutes, wherein sometimes large sample volumes have to be processed. These requirements are not fulfilled by conventional incubation systems. Rather, special aspects of incubators are required.

Furthermore the required sample volumes vary very strongly to yield a desired amount of biomolecules depending on the field of application. Consequently, in practice problems have frequently occurred with apparatus that can only cover a predetermined, narrow volume range in automated processing.

SUMMARY OF THE INVENTION

The apparatus for a purifying of biomolecules comprises a plurality of cavities for accommodating solutions with magnetizable particles. Furthermore the apparatus comprises at least one magnetizable pin that is arranged such that it is insertable in at least one of the cavities, and at least one counter magnet is arranged in at least one bottom area of the cavities.

The lysis cavities 22 and the work plate 80 may be used as cavities within the apparatus 10. Optionally also the yield plate 85 may serve as cavities being arranged outside the apparatus 10 forming a component of the system.

The apparatus and the method according to the invention serve for the completely automated lysis, extraction and purifying of biomolecules (e.g. DNA, RNA or proteins) up to the final eluate containing the biomolecules in an ultrapure suspension, thereby also corresponding to the requirements of clinical in-vitro diagnostics (IVD). The samples are of a highly different nature. The application of the invention reaches from the field of research to the field of IVD. The purifying occurs by adsorption of the biomolecules to magnetizable particles. A complete run of the method according to the present invention is rendered possible within the apparatus. The apparatus executes the complete method for the lysis, extraction and purifying for the corresponding biomolecules without any intervention by laboratory staff. This means that at the start the sample (e.g. blood, patient tissue, blood plasma, blood serum), bearing a machine-readable identification, is placed into the apparatus. Selectively, the sample is attributed to an unambiguously identified processing job. Selectively on the basis of this machine-readable job or based on an input program the apparatus controls the addition of the substances and aids and derives all required process parameters for controlling the method the identification of the sample, the substances and the aids. The complete process is documented and takes place without possibility of interference by the laboratory staff. This means that there is no danger of confusion, a strongly reduced danger of infection, no danger of contamination and cross-contamination.

The present invention is adapted to use a plurality of cavities for accommodating mixtures of samples and reagents. The invention discloses a method for enhancing the minimum and maximum sample volumes and also the amount of processable biomolecules (upscaling).

Moreover, the invention discloses a closable incubation unit adapted to generate a homogeneous temperature within the cavities/in the mixture of samples and reagents in the incubator within a few minutes, and is adapted to stabilize this temperature with great precision without any liquid evaporating.

The invention furthermore discloses a method for lysing and purifying biomolecules within this mixture of samples and reagents using magnet particles or magnetizable particles. This method comprises a mixing of the biomolecules with a binding buffer and magnetizable particles, thereby forming a particle-biomolecule complex, wherein the biomolecules are bound to the magnetizable particles by the binding buffer.

The method furthermore comprises the switching on or inserting of a magnetizable pin. Moreover the method comprises the transporting of the particle-biomolecule complex to a first cavity with a first solution. In addition the method comprises the switching off of the magnetizable pin, an optional switching on of a counter magnet and a mixing of the particle-biomolecule complex with the first solution.

The inventive method and the apparatus for purifying, use two groups of magnets for the purifying of biomolecules after their adsorption to magnetizable particles: a plurality of magnetizable pins and at least one counter magnet. Thereby it is in particular possible to substantially reduce the number of magnetizable “residual particles” present in the eluate; the group of counter magnets allows the retention of the “residual particles” present in the eluate in the cavity. Thereby the purity of the eluate and consequently the functionality of subsequent analyses are improved.

The invention furthermore discloses a system for lysis, extraction and diagnostically purifying of biomolecules. The system according to the invention comprises the apparatus for purifying of biomolecules according to the invention. The system furthermore comprises a control unit for controlling the apparatus. The system yields an eluate with diagnostically purified biomolecules from the sample material. Moreover, a detection unit detects a for detecting a code: The detection unit furthermore transfers the code to a yield cavity.

Furthermore the invention discloses a method for the diagnostically purifying of biomolecules comprising an execution of the method for purifying biomolecules according to the invention to yield an eluate with the diagnostically purified biomolecules from a sample material under the exposure to magnetizable particles.

FIGURES

FIG. 1 a shows a perspective view from above of the apparatus according to the invention

FIG. 1 b shows a perspective view from below of the apparatus according to the invention

FIG. 1 c shows a cross section of the apparatus according to the invention

FIG. 2 shows the flow of the method according to the invention as a flow chart

FIG. 3 a shows an example of a plate with large incubator cavities in 12 rows from above

FIG. 3 b shows an example of a plate with 8 times 12 equaling 96 incubator cavities from above

FIG. 3 c shows a method without upscaling

FIG. 3 d shows a method with an upscaling from a large cavity

FIG. 3 e shows a method with an upscaling from standard cavities

FIG. 3 f shows a method with an upscaling from standard cavities

FIG. 4 a shows a perspective lateral view of an alternative aspect of the apparatus for lysis and extraction

FIG. 4 b shows a perspective view from behind of the alternative aspect of the apparatus for lysis and extraction

FIG. 4 c shows a cross section of the alternative aspect of the apparatus

FIG. 4 d shows a magnetizable pin according to the invention

FIG. 5 a shows a top view of a system according to the invention

FIG. 5 b shows a perspective view of the system according to the invention

FIG. 6 a shows a method for diagnostically purifying biomolecules

FIG. 6 b shows a section of the method for diagnostically purifying biomolecules

FIG. 7 shows a block diagram of the system according to the invention

DETAILED DESCRIPTION OF THE INVENTION

The invention will now described on the basis of the drawings. It will be understood that the embodiments and aspects of the invention described herein are only examples and do not limit the protective scope of the claims in any way. The invention is defined by the claims and their equivalents. It will be also understood that features of one aspect can be combined with features of a different aspect.

FIG. 1 a-c show a perspective view of the apparatus 10 according to one aspect of the invention. The apparatus 10 on one level comprises a row of fasteners or a rack 44 for disposables, e.g. plastic covers 42, an incubation unit 60, a work-area retainer 81 for a work plate 80, an accommodation cavity 70 for the used disposables, as well as a yield-area retainer 86 for a yield cavity 85. A first drive motor 90 a and a first band 92 a are mounted on the level. The work-area retainer 81 and the yield-area retainer 86 each have a plurality of cavities 20.

The apparatus 10 has a frame 45 with magnetizable pins 40 with electric connections 46. The frame 45 comprises a second drive motor 90 b, a second band 92 b and a spindle 92 c.

Below the level, below the work-area retainer 81 counter magnets 50 with electric connections 51 are disposed (see FIG. 1 b). Racks 44 for accommodating a plurality of the disposables 42 are provided next to the incubation unit 60.

The disposables used are preferably plastic covers 42 and consist of for e.g. polypropylene or any other suitable material. For example the aspect of the plastic covers 42 described within the framework of this description is approx. 50.5 mm long and has a diameter of approximately 5.5 mm, widening to approx. 8.5 mm toward the upper end. The wall thickness is approximately 0.5 mm. Of course also different dimensions of the plastic covers 42 are possible. The dimensions of the plastic covers 42 are chosen such that they fit ends 41 of the magnetizable pins 40 well.

Of course also other dimensions of the plastic covers 42 are possible, as long as they fit the ends 41 of the magnetizable pins 40 well and can still dip into the cavities 20 comfortably, without making liquids in the cavities 20 flow over.

For example the magnetizable pins 40 have a bulge 43 (see FIG. 1 c) on which the plastic covers 42 are caught. The plastic covers 42 are disposed in a rack 44 for plastic covers 42 adapted to accommodate plastic covers 42 and is optionally filled by an automatic feeding system (not shown in FIGS. 1 a, 1 b and 1 c).

The incubation unit 60 can accommodate at least one lysis cavity 22 (not shown) in a sample area. The start material or sample material containing biomolecules 28 is placed in the lysis cavity 22. In a fully automated or semi-automated process the filling of the lysis cavity 22 preferably takes place using a suitable pipetting system, which is not shown in FIGS. 1 a and 1 b, as will be explained below in connection with the system according to the present invention (see FIG. 7 a).

Optionally the incubation unit 60 can be closed by a lid 61, as visible in FIG. 4 a, or by a lid 61 with openings 62, as visible in FIG. 1 a. Into these openings 62 in the lid 61 of the incubation unit 60 a suitable pipetting system 840 can be lowered (see FIG. 5 b). The incubation unit 60 allows a heating of the sample area to temperatures above the room temperature. Preferably the openings 62 of the incubation unit 60 are designed such that the openings 62 can be closed by lowering the magnetizable pins 40, which facilitates achieving a target temperature within the lysis cavity 22 inside of the incubation unit 60. Additionally moving the magnetizable pins 40 up and down in the lysis cavity 22 effects a mechanical mixing of the start material of biomolecules 28 and at least one lysis buffer 30 a and added magnetizable particles 35, in order to form a solution 30. The lysis buffer 30 a and the magnetizable particles 35 were filled into the lysis cavity 22 beforehand by the suitable pipetting system 840 (shown on the right in FIG. 5 b).

It is furthermore possible to optionally automatically remove the optional lid of the incubation unit 60 after a selectable time interval, to facilitate the cooling of the solution 30. Additionally or alternatively the cooling process can also be supported by heat-dissipating procedures, e.g. a ventilator.

The work plate 80 can be implemented as a deep-well plate and can contain a first plurality of the cavities 20. For a conventional deep-well plate, each cavity 20 can hold a volume of approx. 2 ml, which is a suitable size to accommodate for example blood samples and to purify biomolecules 28, e.g. DNA or RNA material. This volume allows integrating this apparatus 10 in a fully automated process for purifying biomolecules 28.

The yield cavity 85 can be implemented as a microtiter plate for accommodating purified biomolecules 28 in the solutions 30. Alternatively the yield cavity 85 can also be implemented by one or several lines of individual sample tubes accommodating the purified biomolecules 28.

A rake can be provided to facilitate the slipping off of the disposables above the accommodation cavity 70, for used disposables; in particular the plastic covers 42.

According to a further aspect of the present invention it may be of interest to arrange the accommodation cavity 70 for used disposables between the work plate 80 and the yield cavity 85. It is thus ensured that the disposables used in the purification process do not have to be moved above the yield cavity 85, thereby further reducing unintentional contamination of the purified biomolecules 28.

The frame 45 is moved along one side of the apparatus 10, so that the frame 45 can sweep the rack 44 for plastic covers 42, the incubation unit 60, the work plate 80, the yield cavity 85, the accommodation cavity 70 for used plastic covers 42. In the aspect of the apparatus 10 shown in FIG. 1 a-c the frame 45 is moved along the longitudinal side of the apparatus 10.

The frame 45 is moved by the first actuator 90 a. Preferably the frame 45 is implemented as a sliding carriage on two longitudinal rails 92 and the thrust of the first drive motor 90 a is transferred to the carriage slide via the first band 92 a, so that the frame 45 can be moved along the longitudinal axis of the apparatus 10. The first drive motor 90 a can be controlled by suitable software.

The magnetizable pins 40 can be implemented as permanent magnets. Alternatively it is possible to implement the magnetizable pins 40 as electromagnets. The necessary electric connections 46 for controlling the magnetizable pins 40 implemented as electromagnets are shown in FIG. 1 a.

By means of the second drive motor 90 b the magnetizable pins 40 can be moved on the frame 45 in the z direction. In the embodiment of the apparatus 10 shown in FIG. 1 a-c the movement of the second drive motor 90 b is transferred to the spindle 92 c via the second band 92 b, so that the z position of the magnetizable pins 40 can be varied. The movement of the second drive motor 90 b can also be controlled by software.

A movement of the magnetizable pins 40 along the z direction can be used for mixing the solution 30 within the cavity 20. For a magnetizable pin 40 implemented as a permanent magnet dipping the magnetizable pin 40 into the solution 30 within the cavity 20 generates a magnetic field to which the solution 30 is subjected. In case that the magnetizable pin 40 is implemented as an electromagnet, in addition to lowering the magnetizable pin 40 a current flow of suitable amplitude and direction has to be circuit-switched through the electromagnet, in order to switch the magnetizable pin 40 to magnetic or non-magnetic. Also in this case the moving up and down of the magnetizable pin 40 in the z direction serves for mixing the solution 30 or to improve the collection of magnet particles 35 within the cavity 20.

In the alternative aspect with permanent magnets instead of electromagnets a removal of the magnetizable pin 40 from the solution, corresponds to a switching off of the magnet or of the magnetic field of the magnetizable pins 40. Outside the solution 30 the magnetizable pin 40 does not exert a magnetic field on the solution 30 within the cavity 20.

In the aspect of FIG. 1 a-c the counter magnets 50 are arranged below the work plate 80, near the bottom area 25 of the cavities 20 within the work plate 80. The counter magnets can be identified best in FIGS. 1 b and 1 c. The group of counter magnets 50 comprises at least one counter magnet 50. The counter magnets 50 can be implemented as an arrangement of permanent magnets, so that respectively one of the counter magnets 50 corresponds to one cavity 20 within the work plate 80. The position of the counter magnets 50 can be changed relative to the cavities 20 of the work plate 80. In such an aspect it is possible to approach the cavities 20 with the permanent magnets, so that the cavity 20 is penetrated by the magnetic field of the counter magnets 50 in the bottom area 25.

In a second position of the permanent magnets the counter magnets 50 are sufficiently spaced apart from the cavities 20, so that the magnetic field of the counter magnets 50 essentially vanishes in the bottom area 25 of the cavities 20. The magnetic field of the counter magnets 50 thus can be switched by a suitable position of the counter magnets 50 implemented as permanent magnets. As for the aspect according to FIG. 1 b counter magnets 50 are allocated to the cavities 20. In FIG. 5 b one of the counter magnets 50 is allocated to several of the cavities 20. Likewise it would be possible to allocate to each of the cavities 20 an individual one of the counter magnets 50.

Alternatively it is in addition possible to provide only one row of a plurality of counter magnets 50, so that the number of counter magnets 50 corresponds to the number of columns within the work plate 80. For such an aspect the apparatus 10 comprises a device for moving the row of counter magnets 50 with the movement of the frame 45 and thereby the row of magnetizable pins 40. It would be possible to couple the movement of the row of counter magnets 50 to the movement of the first band 92 a. In the case that the row of counter magnets 50 is embodied as permanent magnets, it is again necessary that the row of counter magnets 50 is mobile in order to switch the magnetic field in the bottom area 25 of the cavities 20 of the work plate 80 either on or off.

Moreover it is possible to implement the counter magnets 50 only as one single counter magnet 50 which is moved from one cavity 20 to the next and moves along a grid in the x and y directions to cover all the cavities 20. However, for a system with a plurality of magnetizable pins 40 such a method is slower than the two above-mentioned aspects of the counter magnets 50.

Independent of the special aspect of the counter magnets 50 these can be implemented as electromagnets which, upon a current flow, induce a magnetic field in the bottom area 25 of the cavities 20. This magnetic field of the counter magnets 50 vanishes substantially as soon as no current flows through the electromagnets any longer. Hysteresis effects within the electromagnets as well as diffusion processes of the magnetizable particles 35 within the solutions 30 limit the speed at which the counter magnets 50 implemented as electromagnets can be switched. The polarity of the current flow is to be chosen in such a fashion that, when the magnetic field is active, the magnetizable particles 35 are subjected to a force in the direction of the bottom area 25 of the cavity 20. For an aspect of the apparatus 10 containing electromagnets as counter magnets 50 it is consequently possible to switch the magnetic field of the counter magnets 50 on and off depending on the current flow. It is of interest to have the software controlling the apparatus 10 to switch the counter magnets 50 on and off. Counter magnets 50 implemented as electromagnets are shown in FIG. 1 b with their electric connections 50 a.

In FIG. 1 c furthermore the solutions 30 within the cavities 20 are shown. The solutions 30 can still contain residual amounts of magnetizable particles 35 even after the last stage of the eluate production. The movement of these magnetizable particles 35 toward the bottom area 25 of the cavities 20 is supported by the counter magnets 50. The effect of the counter magnets generates a shift of the residual magnetizable particles 35 toward the bottom and thereby improves the eluate to be free from particles, which is transferred from the upper layer out of the cavity 20 into the eluate cavity 85 using the pipetting device 840.

FIG. 2 shows a flow of a method 100 according to the invention for purifying biomolecules 28. The apparatus 10 and the method 100 according to the invention allow a complete execution of the method 100 within the apparatus 10.

By way of preparation in a step 102 the frame 45 with the magnetizable pins 40 is moved over the loaded rack 44 for plastic covers 42 and is lowered, so that one plastic cover 42 is slipped onto each magnetizable pin 40.

First the sample to be lysed, contained in a conventional sampling cavity (not shown in FIGS. 1 a and 1 b), plus a buffer are placed 105 in the apparatus 10. The conventional sampling cavity can e.g. be a tube with blood serum, which in addition bears an unambiguous identification. This unambiguous identification of the sample, for example a barcode, rules out any unintentional confusion of the samples. A method 100, which is to be used in the diagnosis of biomolecules 28, has to ensure that several samples can be allocated with absolute certainty and without any danger of confusion. The apparatus 10 and the method 100 according to the present invention rule out such confusions of sample material, since barcode scanners can document the execution of the method 100 in important positions within the apparatus 10.

The sample is aliquoted 110 into the lysis cavity 22 using a suitable pipetting device 840, contained in the apparatus 10 (shown in FIG. 5 b). In addition the suitable pipetting device 840 pipets 115 a lysis buffer 30 a into the lysis cavity 22.

All following examples of buffers are used in the purification procedure for DNA from whole blood; other start materials partly require other buffer systems. An example for such a lysis buffer is Lysis Buffer A for blood samples, such as used in a variety of reaction kits by Invitek. This buffer is based on the patent DE 19856064C2.

A lysis procedure 117 takes place at a possibly elevated temperature provided by the incubation unit 60 accommodating the lysis cavity 22. A mechanical mixing of the solution 30 in the lysis cavity 22 during the lysis procedure 117 is possible by moving the magnetizable pins 40 up and down in the lysis cavity 22. The lysis procedure 117 takes between 10-30 minutes, depending on the biomolecules 28. The mixing during the lysis procedure 117 can selectively take place through cyclical opening of the lid 61 of the incubation unit 60 or also with an open incubation unit 60.

During this lysis procedure 117 the apparatus 10 using the suitable pipetting system 840 prepares cavities 20 arranged in the accommodation area 81 of the work plate 80 with washing buffers 32 a and 32 b and elution buffers 33, preferably first a first washing buffer 32 a, then second a second washing buffer 32 b. Without any limitation more than two washing buffers 32 a, 32 b are possible. Furthermore at least one elution buffer 33 is pipetted. Examples for washing buffers are Wash Buffer I and Wash Buffer II, an example for elution buffers is Elution Buffer D. These buffers are based on the patent DE 19856064 C2.

This is followed by the addition 120 of a binding buffer 30 b and the magnetizable particles 35 to the lysis cavity 22 in suspension. For example Binding Buffer B6 or Binding Buffer. These buffers are based on the patent DE 19856064 C2.

By moving the magnetizable pins 40 of the apparatus 10 up and down, the biomolecules 28, the binding buffer 30 b and the magnetizable particles 35 are mixed 122 within the lysis cavity 22. Optionally also the pipetting device 840 itself can be used for mixing, by absorbing and dispensing the solution. Subsequently the magnetizable pins 40 are switched to magnetic 125 e.g. by switching on the current and the magnetizable particles 35 are collected.

To the magnetizable particles 35 the biomolecules 28 e.g. nucleic acid 28 a have now attached themselves in order to form a particle-biomolecule complex 36. The magnetizable particles 35 with the bound nucleic acid 28 a are transported 130 by the apparatus 10 through the washing buffers 32 a, 32 b; according to the following principle for each washing buffer 32 a or 32 b etc.

The magnetizable pins 40 transport 130 the particle-biomolecule complexes 36 when switched to magnetic and thereby place the bound nucleic acid 28 a in a first cavity 20 a or in a second cavity 20 b with the buffer solution 32 a, 32 b, etc. Subsequently the magnetizable pins 40 are switched off or the permanently magnetic pins 40 are removed 135.

This is optionally followed by a mixing step 140 carried out once or several times, with switching on the counter magnet 50, and the particle-biomolecule complexes 36 shift from the switched-off magnetizable pin 40 to the bottom area 25 of the cavity 20. After concluding the mixing 145 a, 145 b of the solution 30 within the cavities 20 a, 20 b the counter magnet 50 is switched off again 146.

Alternatively the particle-molecule complexes 36 are distributed and mixed 145 a with the washing buffer solution 32 a within the first cavity 20 a or the particle-biomolecule complexes 36 are mixed 145 b with the washing buffer solution 32 b within the second cavity 20 b etc. by moving the magnetizable pins 40 up an down together, of course with the plastic caps or plastic covers 42 disposed on top, so that the pins 40 and the plastic caps 42 dip into the washing buffer solution 32 within the first cavities 20 once or several times. It would furthermore be possible to carry out the mixing 145 a, 145 b of the solution 30 or 32 a, 32 b etc. by alternately switching on and off or moving the magnetizable pins 40 and the counter magnets 50. The mixing 145 a, 145 b of the solution 30 or 32 a, 32 b in this case was carried out by the particle-biomolecule complexes 36 within the solution 30. A movement of the particle-biomolecule complexes 36 between the wall area or bottom area 25 of the first cavity 20 a or the second cavity 20 b and the plastic covers 42 would also effect the mixing 145 a, 145 b of the solution 30. However, this alternative mixing process is limited by the switching cycles of the counter magnets 50 and the magnetizable pins 40.

Switching on 150 the magnetizable pins 40 or inserting the permanently magnetic pins 40 collects the particle-biomolecule complexes 36 from the washing buffer 32 a, 32 b again.

In an inquiry 160 it is determined whether further washing buffers 32 etc. are to be processed. In case of further washing buffers 32 to be processed, this is followed by the transport 130 to the next cavity 20 with the further washing buffer 32.

However, if according to the inquiry 160 no further washing buffers 32 have to be processed, the magnetizable pins 40 and the plastic covers 42 with the particle-biomolecule complexes 36 adhering thereto are lifted from the solution 30 and a waiting step follows to dry 170 the particle-biomolecule complex 36 at least partially. Subsequently the dried particle-biomolecule complex 36 is transported to the cavity 20 with the elution buffer 33 and is placed 180 in the elution buffer 33. Thereupon the magnetizable pins 40 are switched to non-magnetic or the permanent magnets 40 are removed 190 from the cavities 20.

This is optionally followed by switching on 200 the counter magnet 50, whereby the particle-biomolecule complex 36 increasingly shifts from the switched-off magnetizable pin 40 towards the bottom area 25 of the cavity 20 due to the counter magnetic field.

In a subsequent process step a mixing 210 is carried out in order to dissolve the particle-biomolecule complex 36. For this purpose the magnetizable pins 40 and the counter magnet 50 are switched off and the solution 30 with the elution buffer 33 is mechanically mixed in the cavity 20 within the yield cavity 85 by moving the pins 40 up and down in a non-magnetic state (with the magnetic field switched off). Thereby the particle-biomolecule complex 36 is detached, the particles 36 are brought to suspension and the biomolecules 28 a are desorbed from the magnetizable particles 35 (this is the so-called elution).

A subsequent switching on 220 of the magnetizable pins 40 collects the magnetizable particles 35 which are now freed of the biomolecules 28 from the elution buffer 33. The biomolecules 28 or nucleic acids 28 a remain in the elution buffer 33. In addition a certain residual amount of the magnetizable particles 35 can remain in the eluate, which now consists of the elution buffer 33, biomolecules 28 or nucleic acids 28 a and the not completely removed magnetizable particles 35 which disturb the further processing.

A disposal 240 of the magnetizable particles 35 removed upwardly from the elution buffer 33 which still adhere to the magnetizable pins 40 switched to magnetic takes place e.g. into one of the cavities 20 used before with the washing buffer 32 a, 32 b within the work area 80. The disposal 240 is carried out by switching off the magnetic field or removing the permanent magnets. Alternatively the disposal 240 can also take place into the receptacle 70 (the particles remain on the magnetizable pins 40 first).

By now switching on 245 the counter magnet 50 of the cavity 20 with the elution buffer 33 the magnetizable particles 35 remaining in the elution buffer 33 are pulled downward and are thus actively removed from the upper area and are held in the lower area of the cavity 20. Thereby in the upper area of the cavity 20 with the eluate 33 an ultraclean eluate is yielded, containing the biomolecules 28 or the purified nucleic acid 28 a.

By means of the pipetting system 840 this ultraclean eluate is now removed by pipetting 250 the ultraclean eluate into the cavities 20 within the yield cavity 85.

To conclude 270 the method 100 the plastic covers 42 on top of the magnetizable pins 40 can be slipped off at a rake above the receptacle 70. If the magnetizable particles 35 remained on the magnetizable or magnetized pins 40, also the magnetized particles 35 are dropped into the receptacle 70.

The method 100 can purify all types of biomolecules 28 in dependence on the magnetizable particles 35, lysis buffer 30 a, binding buffers 30 b, washing buffers 32 a, 32 b and elution buffers 33. With a suitable combination of the washing buffers 32 (32 a, 32 b, etc.) elution buffer 33 and the individual procedure also combinations of different biomolecules 28 are possible, for example DNA and RNA together, or also separately in two cavities or reaction cavities within the yield cavity 85, or also in two cavities within two separate yield cavities 85.

This combination of the pipetting technology with a magnet separation via magnetizable transport magnets renders possible an unprecedented repertoire of combined purifications and a greater variability concerning the sample amounts to be processed.

The invention contains parallel and serial variants of upscaling a sample amount. Upscaling means a targeted increase of the processable sample volume of a start substance. The parallel and serial variants of the upscaling can also be combined. Upscaling means that within the framework of the upscaling several cavities or partly also greater cavities are used. In the several cavities or the greater cavities the same solution 30 is disposed. The same solution 30 means that substantially the same sample material, the same buffer solutions and the same magnetizable particles 35 are used. In particular the concentrations and/or the substance amounts of the same solutions 30 are substantially equal.

Combining the apparatus 10 and the suitable pipetting system 840 offers the possibility of parallel upscaling. A further aspect of the apparatus 10 according to the invention is designed for the parallel processing of 12 samples, wherein in different configurations also different numbers of samples are possible. The processing path for the sample or the sample material herein is referred to as a channel, thus 12 channels are provided. Several of these purification channels can be used in parallel for one sample, whereby in this example a sample volume can be processed that is up to 12 times greater the initial volume. After the preparation of the eluate with the elution buffer 33 and biomolecules 28 the pipetting system 840 can control the removal by pipetting 250 in such a fashion that the eluate from the several channels is placed in one or several cavities within the yield cavity 85, whereby the biomolecules 28 distributed to several channels are joined again.

Furthermore a serial upscaling can be achieved within the channels if used for the lysis or incubation 117 within the lysis cavity 22 an array with several cavities per channel is used. Therein during the method 100 one channel is used several times with the same sample, in that the start material is distributed to several cavities of the lysis cavity 22 within one channel. Then the particle-biomolecule complexes 36 can be transferred in several partial steps from these cavities of one channel into the cavity 20 within the work plate 80 into the first washing buffer 32 a.

This means that a serial upscaling within one channel can be achieved up to a multiple, e.g. the five-fold sample amount. The objective of this upscaling results from the capacity of the washing buffers, which can be loaded e.g. with up to the five-fold amount of the magnetizable particles 35 from one individual isolation process. Consequently in the case of the combination of the serial upscaling within the channels and the parallel upscaling of several parallel channels the sample volume could, in the extreme case in the example of a 96-cavity microtiter plate, be increased by 96-fold. This means, starting from the usual amount of 0.2 ml, in the extreme case more than 10 ml could be purified. This is the amount which is at most contained in e.g. a standard collection cavity for blood or serum. However, this also means that per run only one sample can be processed. It is an advantage herein that no change in equipment is required, it is e.g. possible to use conventional 96-cavity microtiter plates and uniform plastic covers 42.

FIG. 3 a shows an aspect of a plate with incubator cavities 65. The plate shown in FIG. 3 a with the incubator cavities 65 is a conventional multiwell plate, the cavities of which can accommodate a volume greater than 2 ml. The incubator cavities 65 of the multiwell plate can furthermore be arranged in the apparatus 10 along a working direction of the apparatus 10. This working direction of the apparatus 10 preferably corresponds to the longitudinal direction (x direction) for moving the frame 45.

FIG. 3 b shows an aspect of a lysis cavity 22 which can also be used as a work plate 80. Preferably the lysis cavity and/or the work plate 80 are exchangeable plates, e.g. 96-cavity microtiter plates or deepwell plates (e.g. disposables). Usually such a microtiter plate consists of eight rows (A . . . H) of respectively twelve (1 . . . 12) cavities 65. Of course also different numbers of the rows and columns or also non-orthogonal arrays of cavities 65 or 20 are possible.

The serial upscaling within a channel can take place in the area of the incubation device or the incubation unit 60 within the lysis cavity 22 in several alternative ways. Selectively a special plate is used with particularly voluminous (e.g. more than 10 ml), e.g. elongated incubator cavities 65 (FIG. 3 a). In this case the complete amount of the magnetizable particles 35 is placed into these incubator cavities 65 and is collected from these cavities after adsorbing the biomolecules 28 using magnetic force.

Optionally the transfer of the collected magnetizable particles 35 out of the lysis cavity 22 into the work plate 80 can also take place in several sequential steps. Alternatively a lysis cavity 22 with numerous cavities 20 of a usual size (approx. 1 to 2 ml) can be used e.g. a 96-cavity microtiter plate or a deepwell plate. Then the sample-reagent mixture to be examined is distributed to several cavities 65 of this lysis cavity 22 and the magnetizable particles 35 are first placed into at least one of the cavities. Subsequently, starting with a first filled cavity, the magnetizable particles 35 are sequentially transferred to all cavities filled with this sample-reagent mixture. The magnetizable particles 35 increasingly take up the biomolecules 28 from all these filled cavities. At the end of this procedure the magnetic collection device has collected the biomolecules from all these filled cavities 65 of the lysis cavity 22 and transferred them to a first cavity on the work plate 80.

In a further variant of the serial and parallel upscaling several parallel channels (e.g. 1 . . . 12) of e.g. several cavities 65 each (e.g. A-H) of a lysis cavity 22 are filled with the same sample. The processing steps are then executed one by one, from incubating to extracting up to finalizing the eluate in one respectively last cavity on the work plate 80. From the thus produced last row of cavities with eluate, this eluate can be transferred from all cavities to at least one cavity on the yield cavity 85 using the pipetting device 840.

The invention furthermore provides that within the lysis cavity for mixing the solutions 30, e.g. in a variant with large cavities 20 extending in the longitudinal direction of the apparatus 10, such as the incubator cavities 65 shown in FIG. 3 a, the magnetizable pins 40 are moved not only in the vertical z direction, but additionally along the longitudinally directed x direction of the apparatus 10. The magnetizable pins 40 can also be moved in an oscillating fashion up and down in the vertical z direction and/or back and forth in the longitudinal direction of the apparatus.

FIG. 3 c first shows the process steps without the upscaling. The particle-biomolecule complex 36 is transferred out of the lysis cavity 22 to individual cavities 20 a, 20 b of the work plate, as already described.

FIG. 3 d shows a first variant of the serial upscaling from an incubator cavity 65 of an incubation plate, as shown in FIG. 3 b. The biomolecule complex 36 is collected from the incubator cavity 65 by moving the plastic covers 42 with the magnetizable pin 40 in the x direction and z direction and is transported (130) into the first cavity 20 a of the work plate 80.

FIG. 3 e shows a further variant of the upscaling from a standard plate as lysis cavity 22 into a work plate 80 also with a standard plate. It is possible to move the magnetizable particles 35 from left to right through the rows A, B, C, . . . H. Upon each contact of the particles 35 with the solutions in the lysis cavity 22 a yield of biomolecules 28 is increased, which, forming the particle-biomolecule complex 36, are bound to the magnetizable pins 40 and are thus extracted from the lysis cavity 22. A transfer of the particle-biomolecule complex 36 in this variant takes place in one single transport step 130.

FIG. 3 f shows a further variant of the upscaling from the standard plate as lysis cavity 22 to the work plate 80, which, like in FIG. 3 e, comprises a standard plate. In contrast to FIG. 3 e in FIG. 3 f not all rows of the lysis cavity 22 are passed sequentially. Instead from each of the rows A, B, C, . . . H an individual transport 130 of the particle-biomolecule complexes 36 takes place into the first cavity 20 a of the working plate 80.

The incubation unit 60 in the alternative apparatus in FIG. 4 a to 4 c and 5 a and 5 b is substantially bigger than in FIG. 1 a-1 c, so that the plate with the incubator cavities 65 shown in FIG. 3 a can be accommodated by the incubation unit 60. Furthermore the yield area 86 is no longer arranged on the apparatus 10. The yield area 86 in this aspect forms part of the overall system 800, as will be explained below.

A lid 61 of the incubation unit 60 e.g. is designed in such a fashion that the lid 61 can be opened automatically, e.g. using a hinge. In FIG. 4 a the lid 61 is shown in a closed state. The lid 61 furthermore optionally has small openings 62 for inserting the magnetizable pins 40. The frame 45 can be moved across the incubation unit 60 when the lid 61 is closed and when it is opened, so that the magnetizable pins 40 have access to the incubator cavities 65. On the frame 45 a drip-catcher 46 is arranged, which is best seen in FIGS. 4 a and 4 c. The drip-catcher 46 moves underneath the tips 41 of the magnetizable pins 40, whereby a contamination of cavities 20 by dripping from the magnetizable pins 40 is prevented.

The first drive unit, a for e.g. comprising a motor 90 a and a band 92 a moves the sliding carriage or the frame 45 in the x direction. The second drive unit b, e.g. comprising of a motor 90 b and a band 92 b, opens and closes the lid 61 of the incubation unit 60. A third drive unit c, e.g. with a motor 90 c, moves the arrangement of the magnetizable or permanently magnetic pins 40 and the plastic caps 42 arranged on top of the pins vertically in the z direction. A fourth drive unit 90 d serves to move the counter magnets 50.

For the aspect of the magnetizable pins 40 using electro magnets the third drive unit 90 c for moving the magnetizable pins 40 comprises e.g. a simple motor drive which moves the pins up and down in the z direction. The magnetic field is then switched on and off electrically.

For the aspect of the magnetizable pins 40 using permanently magnetic arrangements the third drive unit 90 c comprises two motors, wherein at least the magnetically effective portion of the permanently magnetic arrangement can be removed from the cavities 20 far enough that this equals switching off the magnetic field.

FIG. 4 d shows a further aspect of the magnetizable pin 40 of the apparatus 10 in cross section. The magnetizable pin 40 comprises a mantle 40 a. On this mantle 40 the plastic cover 42 is disposed approximately in the lower third. The plastic cover 42 provides the tip 41 of the magnetizable pin 40. By changing the plastic cover 42 it is possible in a simple fashion to ensure the cleanliness of the tip 41. The bulge 43 of the magnetizable pins 40 (see FIG. 1 a-c) can be omitted in the further aspect of the magnetizable pin 40.

The further embodiment of the magnetizable pin 40 furthermore comprises a retaining clip 40 g. The retaining clip 40 g serves to hold the plastic cover 44 on the mantle 40 a of the magnetizable pin 40. The retaining clip 40 g connects the plastic cover 44 in a detachable fashion with the mantle 40 a of the magnetizable pin 40. This means that the plastic covers 42 can be taken up by the mantle 40 a and can be shed again reliably.

As an alternative to the detachable connection of the plastic cover 42 to the mantle 40 a of the magnetizable pin using the retaining clip 40 g the dimensions of the mantle 40 a and of the plastic cover 42 can be adjusted to each other in such a fashion that a positive fit of the plastic cover 42 on the cap of the magnetizable pin 40 is achieved.

On the inside of the mantle 40 a of the magnetizable pin 40 a hollow 40 b is disposed. The hollow 40 b is suitable to accommodate a magnetizable element. The hollow 40 b and consequently the magnetizable element can be moved along the longitudinal axis of the magnetizable pin 40 identified by a dashed line. The mobility of the magnetizable element serves to control the effect of the magnetizable element 40 b on the tip 41 of the magnetizable pin 40.

If the magnetizable element 40 b is disposed in the area of the tip 41, the tip 41 is magnetized. This state is also referred to as “switching on of the magnetizable pin 40”. If the magnetizable element 40 b is disposed at a distance from the tip 41, the tip 41 is not magnetized. This state is also referred to as “switching off of the magnetizable pin 40”. By moving the magnetizable element 40 b between the tip 41 and a position at a distance from the tip 41 a change is possible between switching on and switching off the magnetizable pin 40. A speed of this change determines a frequency of the change of a magnetization of the tip 41. The magnetizable element in the hollow 41 can be implemented either as a permanent magnet or as an electromagnet. Electric conducts to the magnetizable element in the hollow 41 and a corresponding voltage supply are required for electromagnets in the hollow 41.

It is furthermore conceivable to combine permanent magnets and electromagnets in the magnetizable pins 40. Thus the electromagnet could be supplied with current in such a fashion that the magnetic field of the electromagnet is opposed to and in total greater than a field of the permanent magnet. Such an aspect would be of interest if a magnetic pulse is to be used to detach the biomolecules 28 from the magnetizable pin 40.

If the magnetizable element 40 b is implemented exclusively as an electromagnet, furthermore the mobility of the magnetizable element in the hollow 40 b relative to the mantle 40 a of the magnetizable pin 40 can be omitted.

The structure of the magnetizable pin 40 shown in FIG. 4 d in particular allows the movement of the mantle 40 a independent of a movement of the hollow 40 b. Consequently, even if a permanent magnet is used as magnetizable element in the hollow 40 b, the magnetizable pins 40 can be lowered into the cavities 20 in a switched-off state.

The apparatus 10 itself has only two axes (x direction longitudinal and z direction vertical) and does not allow any translations in the lateral y direction. However, the suitable pipetting system 840 having at least three axes allows the parallel and serial upscaling as described above.

The invention furthermore provides a system 800 for the so-called “diagnostic purification” of biomolecules 28. The “diagnostic purification” of biomolecules 28 means that the regulations and directives of in-vitro diagnostics (IVD) are fulfilled. The system 800 ensures that no confusion of materials and substances by a user can occur. Moreover all steps carried out by the system 800 are comprehensively recorded.

FIG. 5 a shows a top view of the system 800 according to the present invention. The system 800 comprises the apparatus 10 according to the present invention for purifying biomolecules 28. This apparatus 10 contains the incubation unit 60 and furthermore the already discussed elements of the apparatus 10. The elements of the apparatus 10 are provided with reference numerals in the FIGS. 5 a and 5 b only if relevant for the description of the system 800. Concerning all other elements of the apparatus 10 reference is made to the FIG. 1 a-1 c and FIG. 4 a-4 c.

The system 800 in addition to the apparatus 10 comprises a loading bay 600 for accommodating substances in the system 800. The system 800 furthermore comprises a control unit 860 for controlling the system 800 and the steps carried out by the system 800. The system 800 furthermore comprises a suitable pipetting system or a pipetting unit 840. The suitable pipetting system 840 is shown in a perspective view in FIG. 5 b. The pipetting system 840 can be moved above the system 800 and can sweep the complete base surface of the system 800. The pipetting system 840 is suitable to transfer the substances from the loading bay 600 into the apparatus 10 and/or from the apparatus 10 to the yield area 85. The required pipetting tips are automatically taken up from the storage cavity 880.

In the system 800 the yield area 85 was shifted from the apparatus 10 to an area which is disposed in front of the apparatus 10. This means that the completely purified eluate can be transferred directly into the yield area or into a yield cavity 85 using the pipetting system 840.

The loading bay 600 comprises a number of sample receptacles 610 to accommodate sample cavities 620 (best visible in FIG. 7 b). The sample cavities 620 are identified 1031 using the reading device 500 upon being placed 1030 into the system 800. The reading device 500 for automatically detecting substances placed into the loading bay 600 can for example comprise a barcode reader.

The system 800 comprises a detection device 870 for coded information about used substances. Substances are the samples, liquids, solids in their entirety which are used in the course of the method in the apparatus. The detection comprises the recognition of at least one coding.

In the simplest case only the sample coding is recognized in order to reallocate it in an unaltered and error-free fashion to the result at the end of the procedure.

In the alternative case the detection comprises the recognition of codings of substances, e.g. via barcodes, which can in particular be components of a so-called kit for purifying nucleic acids. In this case a check for consistency can be carried out in order to rule out any confusion within the kit or of the kit itself.

As a further alternative also the identification 1031 of the sample material itself can take place automatically and on the basis of this coding the selection of the method steps to be carried out as well as the selection of substances required for this purpose can be determined via an information system. Through the reading device 500 the system 800 can in particular identify 1031 every sample cavity 620 and a content of the sample cavity 620.

An alternative detection variant can also determine the properties of the used cavities and containers with the cavities disposed thereon, e.g. via a position of the sample cavity 620 in the system 800 a cross section of the sample cavity 620 can be determined. The cross section results from the cross section of the cylindrical opening in which the sample cavity 620 is inserted. A height of the sample cavity 620 and consequently a volume of the sample cavity 620 can be coded on a reading element 501, so that the volume of the sample cavity 620 becomes part of identifying 510 the sample cavity and the sample content.

The loading bay 600 furthermore comprises a row of substance receptacles 650. These consist of cylindrical openings of different sizes, into which the substance cavities 630 (not shown) can be inserted. Upon inserting or pushing in 1035 the substance cavities 630 into the loading bay 600 the position of the substance cavity 630 is determined, and thereby its cross section. Furthermore the content of the substance cavity 620 from the reading element 501 is identified 1036 by the reading device 500. The height of the substance cavity 620 can also be coded on the reading element 501. This is of interest, in particular in the case that substance cavities 620 of different heights can be placed 630 in the loading bay 600.

In the case that very large amounts of samples have to be purified, it is conceivable that the sample material is placed 1035 into the system 800 in one of the substance receptacles 650. The reading device 500 will correctly identify 1031 the inserted sample amount as sample material.

The system 800 furthermore optionally comprises a plurality of sensors 830 to record and log a plurality of parameters 835. Such a parameter can be the temperature of the incubation unit 60 and the dwell time of the sample material in the incubation unit 60, but is not limited thereto. Furthermore a plurality of parameter controls 839 is provided to control the plurality of parameter values in the system 800.

The system 800 is for e.g. suitable to determine required binding buffers 30 b, washing buffers 32 and elution buffers 33 as well as the required magnetizable particles 35 on the basis of the identified sample material via the identification of the substances. Furthermore the system 800 is adapted to determine a required substance amount for each of the elements of the group. Through the identification of the substances it is on the other hand also possible, without interpreting the coding of the sample, to determine the process parameters on the basis of the substances used in the apparatus and to control the actions of the apparatus and the method steps accordingly.

The system 800 can furthermore, on the basis of the identified sample material and/or on the basis of the used reagents and aids, determine a particular IVD-conforming sequence of steps and further process parameters for the diagnostic purification of the biomolecules 28 and for e.g. also the required documentation. Likewise on the basis of the detected information the number of steps required for the diagnostic purification and the selection of further reagents and/or aids can be determined (e.g. the type of magnetizable particles 35).

FIG. 5 b shows a perspective view of the system 800 according to the invention. In this representation the sample cavities 620 are recognizable particularly well.

FIG. 6 a shows an extension of the method 100 carried out as a method 900 for the diagnostic purification of biomolecules 28. For the method 900 the method 100 of FIG. 2 is extended. More specifically, FIG. 8 a shows an extension of the step 102 for taking up the plastic covers 42 on the magnetizable pins 40 in the method 100. If the method 100 is to be extended in the form of the method 900, step 102 has to be expanded.

In a step 1005 first a plate with incubator cavities 65 is loaded into the incubation unit 60. Subsequently in a step 1010 first a work plate 80 is loaded into the work-plate accommodation 81 of the apparatus 10.

This is followed by an enquiry 1012 to check whether plastic covers 42 have to be refilled in the racks 44. In the case that a refill is necessary, the racks 44 are refilled in a step 1015.

Subsequently in a step 1020 the work plate 80 is loaded with the required elution solutions and/or the washing buffers. Finally it is checked in an inquiry 1021 whether the receptacle 70 for the used plastic covers 42 has to be emptied. If required, the emptying is carried out with step 1025.

In a step 1030 the sample cavities are placed into the loading bay 600. Therein the identification of the inserted sample cavities takes place in step 1031. Subsequently in a step 1035 the substance cavities are placed into the loading bay 600, whereupon the inserted substance cavities are identified in step 1036.

On the basis of the identified sample material and/or the identified substances the system 800 determines in step 1040 a suitable multiwell plate and loads this into the incubation unit 60. Therein a required incubation volume is taken into account.

This is followed by step 1045, where the information read by the reading device 500 is recorded and transmitted to the control unit 860 and/or the documentation unit 850. Furthermore step 1045 allows a consistency check of the identified substances in the loading bay 600. Thereby for e.g. the use of a false buffer solution with a sample material can be prevented, so that falsely positive or falsely negative results of the purification can be prevented.

This is followed by a step 1050. In the step 1050 required process parameters 850 are determined.

FIG. 6 b shows the individual stages of the step 1050. First in a step 1055 the required substances are determined among the identified substances in the loading bay 600. Subsequently in a step 1060 a respective substance amount is determined for each of the required substances to carry out the method 900. The substances involved in the method 900 are at least one element of the substances discussed above.

In a step 1065 the required parameter values 835 are determined on the basis of the identified substances. This is followed by step 1070 in which the required process steps are determined on the basis of the identified substances.

Subsequently in step 1075 the apparatus 10 is controlled by the control unit 860 of the system 800. The control unit 860 monitors the apparatus 10 during the execution of the method 100. Step 1075 also comprises the monitoring of the parameter values 835 during the execution of the method 100.

If the method 900 is carried out, the step 105 (FIG. 2) for supplying the sample can be omitted, since the sample material was already supplied in step 1030 (FIG. 6 a).

FIG. 7 shows a block diagram of the system 800 according to the invention. The system 800 comprises a parameter control 839 to control a plurality of parameter values 835. The parameter values 835 are detected by a plurality of sensors 830. Furthermore the system 800 comprises the apparatus 10 according to the invention, a pipetting device 840 as well as a documentation unit 850 for documenting detected information, for example by the reading device 500. The information read by the reading device 500 is recorded in a recording module 870 and forwarded to the system control 860 and/or the documentation unit 820. The control unit 860 controls and regulates the system 800 on the basis of programs and the recognized information about the substances. The elements of the system 800 shown in FIG. 7 are connected to each other and communicate with each other.

The system 800 comprises software for controlling the method shown in FIGS. 8 a and b. This software is run by a microprocessor and can be programmed in any suitable programming language. 

1.-34. (canceled)
 35. A system for diagnostically purifying biomolecules comprising: a plurality of cavities for accommodating solutions with magnetizable particles, at least one magnetizable pin arranged in such a fashion that it can be inserted in at least one of the cavities, a control unit for controlling the system, wherein the system yields an eluate with diagnostically purified biomolecules from the sample material, and a detection unit for recognizing a coding and for transferring the coding to a yield cavity after the diagnostically purifying.
 36. The system according to claim 35 furthermore comprising: a loading bay for accommodating substances, wherein the substances are at least one of a sample material, a buffer solution or magnetizable particles, a reading module for automatically identifying the substances and the sample material upon accommodating in the loading bay, an incubation unit for heating, thermostating and incubating the samples in a lysis cavity, a pipetting unit for transferring the substances, a receptacle, and at least one yield cavity.
 37. The system according to claim 35, furthermore comprising: apparatus for improving the quality of the eluate before removing by pipetting adapted to keep the magnetizable particles away from a tip of the pipetting device.
 38. The system according to claim 35, furthermore comprising: sensors for recording and logging a plurality of parameters using a documentation unit.
 39. The system according to claim 35, furthermore comprising: a plurality of parameter controls for controlling the plurality of parameters using the control unit.
 40. The system according to 36, wherein the control unit is adapted to check a consistency of the substances disposed in the loading bay based on the substances in the loading bay.
 41. The system according to claim 40, wherein the control unit is adapted to check, if the consistency of the substances disposed in the loading bay is correct based on a composition of a kit.
 42. The system according to claim 40, wherein the control unit is adapted to output a message in the case that the consistency of the substances in the loading bay is not correct.
 43. The system according to claim 40, wherein the control unit is adapted to prevent an opening of the loading bay during the controlling of the system, in case that the consistency of the substances disposed in the loading bay is correct.
 44. The system according to claim 36, wherein the system is adapted to determine a substance amount of required buffer solutions in the loading bay based on the identified substance in the loading bay.
 45. The system according to claim 36, wherein the system is adapted to determine a substance amount of the required magnetizable particles based on the identified substance in the loading bay.
 46. The system according to claim 36, wherein the system is adapted such that the system determines a number and a sequence of process steps of a method for diagnostically purifying biomolecules based on the identified substance in the loading bay.
 47. The system according to claim 36, wherein the system is adapted to determine an appropriate incubation volume for incubating based on the identified substance in the loading bay.
 48. The system according to claim 47, wherein the system is adapted to determine a suitable multiwell plate for incubating based on the appropriate incubation volume.
 49. The system according to claim 35, wherein the system is adapted to automatically select, request and use a suitable cavity from the group consisting of lysis cavity, incubator cavity, work plate, cavity of the work plate, yield plate.
 50. A method for diagnostically purifying biomolecules comprising: mixing the biomolecules with a binding buffer and magnetizable particles thus forming a particle-biomolecule complex, wherein the biomolecules are bound by the binding buffer to the magnetizable particles, switching on at least one magnetizable pin, transporting the particle-biomolecule complex to a first cavity with a first solution, switching off the magnetizable pin, mixing the particle-biomolecule complex with the first solution; for yielding an eluate with the diagnostically purified biomolecules from a sample material subjected to the effect of magnetizable particles.
 51. The method according to claim 50, wherein the method furthermore comprises: inserting at least one sample cavity with the sample material in the loading bay, identifying the sample material upon accommodation in the loading bay, inserting at least one substance cavity with a substance in the loading bay, identifying at least one substance upon accommodation in the loading bay, checking a consistency of the substances in the loading bay with the sample material, transferring the substances and/or the sample material.
 52. The method according to claim 50, the method furthermore comprising: recording a plurality of parameters, controlling the plurality of parameters.
 53. The method according to claim 50, furthermore comprising: determining a plurality of process parameters based on the at least one identified substance.
 54. The method according to claim 53, wherein the determining of the plurality of process parameters based on the at least one identified substance comprises: determining at least one buffer solution, determining one required magnetizable particle, determining one required elution solution.
 55. The method according to claim 50, furthermore comprising determining one substance amount of the substances, determining required parameter values for executing the method, determining a number and a sequence of process steps of the method for diagnostically purifying biomolecules, determining a suitable incubation volume, determining a suitable multiwell plate for incubating based on the suitable incubation volume.
 56. The method according to claim 55, wherein determining the substance amount of the substances comprises: determining one substance amount of at least one buffer solution, determining one substance amount of one magnetizable particle, determining one suitable elution solution. 